瘦蛋白对犊牛肝细胞PC mRNA表达及其活性的影响

Effects of Leptin on Pyruvate Carboxylase mRNA Expression and Activity in Vitro Culture Bovine Hepatocyte

取单层培养72 h生长良好的犊牛肝细胞,采用单因素重复试验,分别添加0,2.5,5.0,10.0,50.0,100.0 ng/mL的牛重组牛瘦蛋白(Leptin),每个处理3个重复(每重复2孔)。再培养12 h后分别提取RNA并制备细胞上清液。应用荧光定量PCR方法检测外源leptin对肝细胞糖异生关键酶丙酮酸羧化酶(Pyruvate car-boxylase,PC)基因表达的影响,同时用比色法检测其对肝细胞PC活性的影响。结果表明:一定浓度的Leptin显著抑制了肝细胞PC mRNA表达,降低了PC活性。

A single factor duplicate test was designed to investigate whether leptin affects the expression of pyruvate carboxylase（PC） mRNA and the activity of PC in vitro culture bovine hepatocyte.Leptin was added to the media with 0,2.5,5.0,10.0,50.0 and 100.0 ng/mL.Abundance of PC mRNA in vitro culture bovine hepatocyte cultured for 12 hours after leptin was added was determined by real-time fluorescence quantitative RT-PCR,and activity of PC was determined by colourimetry.The results showed that the leptin could restrain the expression of PC mRNA and the activity of PEPCK in vitro culture bovine hepatocyte.